Figure 1
From: Muscarinic receptor agonist-induced βPix binding to β-catenin promotes colon neoplasia
Colocalization of β-catenin and βPix in the nuclei of human colon cancer cells that overexpress M3R. (a) Overexpression of M3R, but not M1R, was observed in colon cancer stem cells. Images show H&E (left panels) and immunofluorescent staining with DAPI (nuclear stain, blue), CD133 (red-Alexa Fluor 594), M1R, and M3R (both green-Alexa Fluor 488), and merged images. Dashed boxes in H&Es show areas from which images were enlarged. CD133 staining reveals scattered colon cancer stem cells (white arrow) and colon cancer stem cell clusters around blood vessels (invasive front; yellow arrows). Top panels: M1R overexpression was not detected in colon cancer stem cells. Bottom panels: M3R overexpression was detected in colon cancer stem cells. Size bars in H&E images are 50 µm for M1R staining and 100 µm for M3R staining. (b) β-catenin and βPix colocalize in the nucleus of invasive human colon adenocarcinoma cells. Normal colon (top) and poorly differentiated cancer (bottom) tissues obtained at surgery from the same person were stained with H&E and DAPI, and immunostained with anti-β-catenin and anti-βPix antibodies. In normal colon and the cancer core, β-catenin and βPix staining was primarily membranous (arrowheads). In contrast, in cells at the invasive front (delineated by dashed lines in the H&E and merged images), β-catenin and βPix were co-localized to dysplastic nuclei (arrows). Size bars are 20 μm except for 5 μm in the high magnification (High Mag) images. Images are representative of n = 6 cancers.
